Bacanelli Gisele, Araujo Flabio Ribeiro, Verbisck Newton Valerio
Biotechnology and Biodiversity of the Central Western Region Postgraduate Program, Federal University of Mato Grosso do Sul, Campo Grande 79070-900, MS, Brazil.
Embrapa Beef Cattle, Av. Rádio Maia, 830, Campo Grande 79106-550, MS, Brazil.
Microorganisms. 2023 Jun 29;11(7):1692. doi: 10.3390/microorganisms11071692.
is the microorganism that causes tuberculosis, a disease affecting millions of people worldwide. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a fast, reliable, and cost-effective method for microorganism identification which has been used for the identification of spp. isolates. However, the mycobacteria cell wall is rich in lipids, which makes it difficult to obtain proteins for MALDI-TOF MS analysis. In this study, two cell preparation protocols were compared: the MycoEx, recommended by MALDI-TOF instrument manufacturer Bruker Daltonics, and the MycoLyser protocol described herein, which used the MagNA Lyser instrument to enhance cell disruption with ethanol. Cell disruption and protein extraction steps with the two protocols were performed using the H37Rv strain, and the MALDI-TOF MS results were compared. The MycoLyser protocol allowed for improved Biotyper identification of since the log(score) values obtained with this protocol were mostly ≥ 1.800 and significantly higher than that underwent MycoEx processing. The identification reliability was increased as well, considering the Bruker criteria. In view of these results, it is concluded that the MycoLyser protocol for mycobacterial cell disruption and protein extraction improves the MALDI-TOF MS method's efficacy for identification.
是导致结核病的微生物，结核病影响着全球数百万人。基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）是一种快速、可靠且经济高效的微生物鉴定方法，已用于 菌株分离物的鉴定。然而，分枝杆菌细胞壁富含脂质，这使得难以获得用于MALDI-TOF MS分析的蛋白质。在本研究中，比较了两种细胞制备方案：MALDI-TOF仪器制造商布鲁克道尔顿公司推荐的MycoEx方案，以及本文所述的MycoLyser方案，该方案使用MagNA Lyser仪器通过乙醇增强细胞破碎。使用H37Rv菌株对两种方案的细胞破碎和蛋白质提取步骤进行操作，并比较MALDI-TOF MS结果。MycoLyser方案可改善 的Biotyper鉴定，因为使用该方案获得的log(score)值大多≥1.800，且显著高于采用MycoEx处理的结果。根据布鲁克标准，鉴定可靠性也有所提高。鉴于这些结果，得出结论：用于分枝杆菌细胞破碎和蛋白质提取的MycoLyser方案提高了MALDI-TOF MS方法对 鉴定的功效。
